Fumiko Satoh

Immunohistochemical analysis of GCDFP-15 and GCDFP-24 in mammary and non-mammary tissue

BackgroundGross cystic disease fluid protein (GCDFP)-15, a major constituent protein in breast cysts, is known to be a marker of breast cancer, while the diagnostic value of GCDFP-24, a protein with a molecular weight of 24 000 daltons, has not been determined. The aim of this study was to elucidate the usefulness of GCDFP-24 for the differential diagnosis of breast cancer in combination with GCDFP-15 and to characterize the histologic features of GCDFP-24-positive breast cancer.MethodsA total of 326 samples including non-neoplastic tissue and benign and malignant tumors from several anatomic sites were examined using commercially available monoclonal antibodies against GCDFP-15 and GCDFP-24.ResultsIn non-mammary tissue, GCDFP-15 was detected in skin, salivary gland, bronchial gland, prostate and seminal vesicle, and GCDFP-24 was detected in apocrine glands and peripheral nerve. Thirty-seven (44.6%) and 22 (26.5%) samples of 83 breast cancers were positive for GCDFP-15 and-24, respectively. Combined assays of GCDFP-15 and -24 raised the positive rate to 50.6%. The markers were not detected in tumors originating from gastrointestinal tract, bronchopulmonary structures or the genitourinary system. Breast cancers positive for both GCDFP-15 and GCDFP-24 were of lower histologic grade according to Bloom & Richardson’s scoring system (p<0.05).ConclusionImmunohistochemical analysis of GCDFP-24 in combination with GCDFP-15 expression was useful for definitive diagnosis of breast cancers, and the expression of these markers correlated with low grade breast cancer.

Comparative immunohistochemical analysis of developing kidneys, nephroblastomas and related tumors: considerations on their histogenesis.

Immunoperoxidase analysis was performed to evaluate the phenotypic expression of eight renal differentiation antigens in five nephroblastomas, one clear cell sarcoma of the kidney (CCSK), one rhabdoid tumor of the kidney (RTK), and four related tumors. A total of 19 fetal and pediatric kidneys, including two 6th‐week mesonephric tissues, were comparatively studied. All the specimens were fixed in formalin and embedded in paraffin. Neural cell adhesion molecule (NCAM), a marker of the nephrogenic zone of the developing kidney, was consistently expressed in the epithelial and blastematous components of nephroblastomas of the common type. The epithelial components also commonly expressed NK1 and Leu 7 (CD57), and the findings may reflect that both were positive in immature proximal tubules directly differentiating from the NCAM‐positive immature fetal tubuloglomerular buds. In two cases, the epithelial component was immunoreactive for CD10 and WT1 gene product (WT1‐GP). Leu M1, epithelial membrane antigen and CA15–3 were only focally expressed in nephroblastomas. Rhabdomyoblasts in the stroma were positive for WT1‐GP. CCSK was featured by the expression of NCAM and CD10. In RTK, focal epithelial differentiation was discerned, with focal positivity of WT1‐GP and negativity of NCAM. In congenital mesoblastic nephroma, the stromal spindle cells were strongly immunoreactive for WT1‐GP, while WT1‐GP was not expressed in solitary multilocular cyst of the kidney. Pancortical nephroblastomatosis was featured by the diffuse subcapsular reappearance of immature metanephric tissue. Nephroblastomas and related conditions thus offer an adequate model for studying human nephrogenesis.

“dead in Hot Bathtub” Phenomenon: Accidental Drowning or Natural Disease?

AbstractSudden death in a hot bathtub occurs frequently in Japan, particularly among elderly people. This retrospective report describes the epidemiologic circumstances and physical findings at autopsy. In total, 268 victims were found unconscious or dead during tub bathing. After postmortem examination, the manner of death was judged as natural cause in 191 (71.2%) and accidental drowning in 63 (23.5%) cases. Mean age (SD) was 72.1 (15.2) years with no significant difference between males and females. A seasonal difference was evident: the winter displayed the highest frequency. Drowning water inhalation, which was confirmed in 72% of victims, was absent in the others. The most common observations on postmortem examination were cardiac ischemic changes and cardiomegaly. Water inhalation signs were evident in a significantly fewer victims exhibiting these factors. In contrast, inhalational findings were observed more frequently in victims with other backgrounds such as alcohol intake, mobility disturbance, and history of epilepsy. Annual mortality in Japan from accidental drowning in persons aged older than 75 years is 33 deaths per 100,000 population. However, this number may be considerably underestimated as pathologists tend to regard lack of water inhalation as indicating a natural cause of death. Confusion in diagnosis remains consequent to the accidental and natural aspects of “dead in hot bathtub” phenomenon.

Fine needle aspiration cytology of glycogen-rich clear cell carcinoma of the breast : A case report

BACKGROUND Glycogen-rich clear cell carcinoma (GRCC) is a rare variant of breast carcinoma that was first described by Hull et al in 1981. The histologic findings have been described; however, the cytologic characteristics have not been well described. CASE A 44-year-old female exhibited a palpable mass in her right breast. The fine needle aspirate contained cell clusters in a necrotic background. In addition to carcinoma cells with increased degrees of nuclear atypia, carcinoma cells with less atypia were identified. The tumor cells contained small round or ovoid nuclei and abundant, clear cytoplasm. The tumor, resected by modified radical mastectomy, was diagnosed as GRCC with detection of glycogen accumulation in the cytoplasm. CONCLUSION Although rare, glycogen-rich clear cell carcinoma should be considered in the differential diagnosis of breast cancer in fine needle aspiration cytology to prevent misdiagnosis because of its varied morphologic patterns and less atypical cytologic features.

MicroRNA Stability in Postmortem FFPE Tissues: Quantitative Analysis Using Autoptic Samples from Acute Myocardial Infarction Patients.

MicroRNAs (miRNAs) are very short (18–24 nucleotides) nucleic acids that are expressed in a number of biological tissues and have been shown to be more resistant to extreme temperatures and pH compared to longer RNA molecules, like mRNAs. As miRNAs contribute to diverse biological process and respond to various kinds of cellular stress, their utility as diagnostic biomarkers and/or therapeutic targets has recently been explored. Here, we have evaluated the usefulness of miRNA quantification during postmortem examination of cardiac tissue from acute myocardial infarction (AMI) patients. Cardiac tissue was collected within one week of the patient’s death and either frozen (19 samples) or fixed in formalin for up to three years (36 samples). RNA integrity was evaluated with an electropherogram, and it appears that longer RNAs are fragmented after death in the long-term fixed samples. Quantitative PCR was also performed for seven miRNAs and three other small RNAs in order to determine the appropriate controls for our postmortem analysis. Our data indicate that miR-191 and miR-26b are more suitable than the other types of small RNA molecules as they are stably detected after death and long-term fixation. Further, we also applied our quantitation method, using these endogenous controls, to evaluate the expression of three previously identified miRNA biomarkers, miR-1, miR-208b, and miR-499a, in formalin-fixed tissues from AMI patients. Although miR-1 and miR-208b decreased (1.4-fold) and increased (1.2-fold), respectively, in the AMI samples compared to the controls, the significance of these changes was limited by our sample size. In contrast, the relative level of miR-499a was significantly decreased in the AMI samples (2.1-fold). This study highlights the stability of miRNAs after death and long-term fixation, validating their use as reliable biomarkers for AMI during postmortem examination.

SNP association and sequence analysis of the NOS1AP gene in SIDS

One of the speculated causes for sudden infant death syndrome (SIDS) is hereditary disease, in which long QT in electrocardiogram has been investigated in the view of mutations in various ion channel genes. In the present study, a novel QT interval determinant of SNP (rs10494366) in NOS1AP is genotyped in SIDS subjects (n=42) and the control group (n=210). Subjects carrying TT genotype was significantly associated with SIDS, compared with those carrying the TG of GG genotype (95% confidence interval 1.28-8.45). Sequence analysis revealed that one non-synonymous substitution in exon 8 (rs12817159) was observed in one subject, in addition to six common SNPs in exons and introns. This postmortem association study showed variations in NOS1AP might be involved in occurrence of SIDS.

Sarcomatoid carcinoma of the renal pelvis: A case report

A case of sarcomatoid carcinoma of the renal pelvis is reported. A 72‐year‐old male was admitted with a chief complaint of right flank pain. Clinical imaging studies revealed marked dilation of the right renal pelvis caused by a tumor at the pyeloureteric region. Right nephrectomy was performed in June 1999. The 8.4 × 6.5 cm tumor was grossly polypoid in appearance and protruded into the renal pelvis. Histologically, sarcomatoid spindle cells predominated over the carcinomatous component (mainly transitional cell carcinoma, partly associated with squamous cell and adenocarcinoma components). Osteoclast‐like CD68‐positive multinucleated giant cells were scattered. The sarcomatoid component was immunoreactive for both cytokeratin and vimentin. Sarcomatoid cells negative for cytokeratin were also noted. In both the sarcomatoid and carcinomatous components, nuclear overexpression of p53 oncoprotein was confirmed. The histogenesis of sarcomatoid carcinoma of the renal pelvis is discussed.

Acute death due to hyperextension injury of the cervical spine caused by falling and slipping onto the face

This retrospective study presents findings of cases involving fatal injuries, in which the victim was found dead at the scene, resulting from cervical hyperextension force attributable to a fall from a low height. External postmortem examination of 14 victims revealed that abrasions and lacerations of the face or the forehead are typical indicators of a direct impact. Either a disruption at the disk space or a transverse fracture of the vertebral body was apparent in the spinal column. The most frequent disk disruption injury occurred at the inter-vertebral space between C4 and C5, and double disruptions were observed in four instances. The damaged cord demonstrated central hemorrhage; moreover, axonal fragmentation and neuronal chromatolysis in the white matter column were evident histopathologically. The elderly victims (mean age, 64.7 years), many of whom displayed elevated blood alcohol levels, experienced the injury consequent to a fall from a low height, a fall during bicycling or slipping on a slope under accidental circumstances.

Sudden death caused by chronic Chagas disease in a non-endemic country: Autopsy report

Chagas disease is a tropical disease that is prevalent in Latin America. Described herein is an autopsy case of the sudden death of a 48‐year‐old Brazilian man who had stayed in Japan for 7 years. The man, who had a history of Chagas disease, collapsed unexpectedly at work. Because the cause of death was unknown, forensic autopsy examination was performed. As gross findings, the heart was dilated and rounded with an increase in size and weight. The esophagus and large intestine were dilated moderately, with extensive interstitial inflammatory infiltration in the cardiac muscle, but no apparent parasite nest was observed in various tissues. On post‐mortem laboratory examinations, indirect immunofluorescence antibody test indicated the presence of IgG antibody specific to Trypanosoma cruzi in the serum. Subsequent polymerase chain reaction amplification using DNA extracted from blood yielded the specific product derived from T. cruzi genomic DNA. These examinations indicate that the infection had resulted from the Tripanosoma parasite. The cause of death was judged to be chronic cardiomyopathy caused by Chagas disease. It is important for pathologists to know the possible involvement of chronic Chagas disease in sudden unexpected deaths in the current globalized society of Japan.

Evaluation of the allele-sharing approach, known as the IBS method, in kinship analysis.

To infer relatedness from genetic data based on short tandem repeats, the exact method, in which shared allele frequencies are applied to relevant equations, has been conventionally used. An alternative approach is the IBS method that is based on the number of shared alleles between individuals. In the present study, the performance of the IBS method in pairwise kinship analysis was compared with the exact method using simulated data of 10,000 genotype pairs for 15 loci in the ABI Identifiler system. The likelihood ratio in allele-sharing of zero, one and two was calculated from joint probabilities based on allele frequencies of the Japanese population. Whereas the IBS method generally produced lower values of combined indices, smaller deviations of the distributions were evident. The threshold for identification of full siblings relative to non-relatives was comparable with that of the exact method, indicating that both inference powers were almost identical. The likelihood ratio in the IBS method depends on the heterozygosity at a locus, and heterozygosities of the 15 loci were consistent across various population groups, particularly in East Asians. The convenience of fixed LR values in the IBS method is beneficial for cases with uncertain allele frequencies and rare alleles.